Viral fibers

ABSTRACT

Long rod shaped M13 viruses were used to fabricate one dimensional (1D) micro- and nanosized diameter fibers by mimic the spinning process of the silk spider. Liquid crystalline virus suspensions were extruded through the micrometer diameter capillary tubes in cross-linking solution (glutaraldehyde). Resulting fibers were tens of micrometers in diameter depending on the inner diameter of the capillary tip. AFM image verified that molecular long axis of the virus fibers were parallel to the fiber long axis. Although aqueous M13 virus suspension could not be spun by electrospinning, M13 viruses suspended in 1,1,1,3,3,3-hexafluoro-2-propanol were spun into fibers. After blending with highly water soluble polymer, polyvinyl 2-pyrolidone (PVP), M13 viruses was spun into continuous uniform virus blended PVP (virus-PVP) fibers. Resulting virus-PVP electrospun fibers showed intact infecting ability to bacterial hosts after suspending in the buffer solution.

CROSS-REFERENCE TO RELATED PATENT APPLICATIONS

This application claims benefit to provisional application 60/510,862filed Oct. 15, 2003 to Belcher et al., which is hereby incorporated byreference in its entirety.

This invention was developed under the following grants from the federalgovernment: Grant No. DAAD19-03-1-0088, from the Army Research Office.The government has certain rights in the invention.

INTRODUCTION

The introduction section provides reference to a number of technicalpublications which are found at the end of the specification and can beused as a guide to one skilled in the art to practice the presentinvention. No admission is made that any of these references are indeedprior art.

Efforts to mimic the unique structures and specific functions of naturalsystems have provided various useful tools and materials in nanoscience[1-14]. Biosystems produce highly programmed, self-assembled,self-templated structures [6-11]. For example, a small percentage ofprotein in abalone shell nucleates a CaCO3 protein composite that is3000 times tougher than pure CaCO3 [1]. By mimicking thebiomineralization process in nature, it has been shown that proteinsequences, selected through fast evolution of a genetically engineeredvirus library on the bench top, can specifically bind to and nucleatedesired materials [3-7]. The one-pot synthetic route provided by thesegenetically programmed viruses results in self-assembled, highly orderednanocrystal composite materials [4].

The unique properties of biological materials are not limited to theirstructures and functions. Silk spiders and silk worms spin highlyengineered continuous fibers by passing aqueous liquid crystallineprotein (fibroin) solution through their spinneret [15-18]. Once thefibroin solution is released to the air, it hardens into a flexible andhighly oriented semicrystalline fiber that is stronger than any otherpolymer fiber spun [17].

Recently, much research has been focused on producing nanosizedone-dimensional materials such as nanorods, nanowires, and nanofibers[19-24], as well as two- and three-dimensional materials. Among them,nanofiber fabrication using electrospinning has been an efficient meansof generating high surface-to-volume ratios of materials that maypossibly be used as highly sensitive sensors, functional membranes,tissue repair applications, and mechanical structures. Electrospinningis a process of using high electric fields to make narrow fibers withdiameters ranging from tens of nanometers to micrometers [23-27]. Whenan electric field is applied to suspensions extruded through a narrowspinneret, an electrostatically charged hemispherical liquid surface atthe tip of the spinneret is elongated and forms a conical surface knownas Taylor cone [27]. As the electric field is increased to a criticalvalue, the suspension is violently ejected from the conical surface tothe grounded collecting plates. The ejected fibers randomly separate andquickly dry to form narrow diameter fibers. Depending on parameters,such as the electric field applied, distance between tip and collector,and viscosity and conductivity of the suspension, micro- or nanometerscale fibers can be fabricated. The distance between the tip andgrounded collecting plate is roughly inversely proportional to thediameter of the fibers because greater chances of fiber separation existduring flight [28,29]. Concentration, which directly affects theviscosity of the suspension, can play an important role inelectrospinning. Below a particular concentration, suspensions at timesare electrosprayed and not electrospun, resulting in deposited dropletsof suspension. As this concentration is approached, bead and stringshaped fibers are formed [29-30]. Increasing the concentration resultsin the formation of continuous fibers. However, above an upperconcentration limit, the solution at times becomes too viscous to bepulled by electrostatic forces. The continuous fibers can be convertedinto non-woven fabrics, which may be useful for synthesizing novelmembranes due to their high surface-to-volume ratios.

A need exists to fabricate wet spun or electrospun fibers usingnanometer scale liquid crystalline viral particle suspensions where theparticle composition and function can be easily modified genetically toinclude binding or nucleating conjugate moieties for inorganic and/ororganic materials, while maintaining the fibrous structure end product.If the conjugate materials are inorganic, this provides fabrics withcontrolled electrical, magnetic, or mechanical properties. If theconjugate particles have one or more binding functions, thefibers/fabrics could be used as filtration detectors for airborne ordissolved analytes. Combinations of material composition andnanoparticle function could be employed for more complex applications ofdetection or unique flexible devices.

SUMMARY

The invention provides a plurality of methods of fabricating virus-basedmicro- and nanometer scale fibers that, generally, can be in broad termsa mimic of the spinning process of silk spiders. In some embodiments ofthe invention, wet spinning and electrospinning were used to fabricatevirus- based micro- and nanofibers, respectively. In some embodiments,the resulting fibers showed nematic ordered morphologies due to flowforces. In additional embodiments, the virus was blended with asynthetic carrier polymer, poly(vinyl pyrrolidone) (PVP), to improve itsprocessing ability. The resulting virus blended PVP fibers werecontinuous and could be formed into non-woven fabrics mats that retaintheir ability to infect bacterial hosts.

In particular, the present invention provides a method of forming aviral fibrous material comprising a plurality of fibers comprising thesteps of providing virus particles and a solvent in a fiber spinningcomposition, and then fiber spinning the virus particles to form thefibrous material. The spinning can be a wet fiber spinning, anelectrospinning, a microfiber spinning, or a nanofiber spinning. Thefiber spinning composition can be a lyotropic liquid crystallinecomposition. The virus particles can be bacteriophage virus particles.The virus particles can form lyotropic liquid crystalline solutions. Thevirus particles can possess specific binding regions on their particlesurface. The virus particle can possess selective binding regions ontheir surface and can be bound to a conjugate material. The conjugatematerial can be an organic material, a particulate material, or anucleated nanoparticulates material. The virus particles can possessgenetically engineered expressed peptide sequences for selective bindingto a conjugate material. Fiber spinning can be carried out withcrosslinking of the viral particles to form cross-linked fiber. Afterspinning, the fibrous material can be liquid crystalline in the semi-dryor dried, solid state. The material can be birefringent and show thenematic phase. The virus particles can possess genetically engineeredexpressed peptide sequences for specific binding to or nucleation ofconjugate material, wherein the fibrous material comprises virus nucleicacids which can be harvested and amplified.

Moreover, the present invention provides a method of forming agenetically engineered fibrous material comprising a plurality of fiberscomprising the step of fiber spinning genetically engineered virusparticles to form the fibrous material, wherein the virus particles ofthe fibrous material are specifically bound to a conjugate materialafter fiber spinning or are capable of specifically binding to aconjugate material after fiber spinning, and the virus particles of thefibrous material retain the virus structure after fiber spinning. Thevirus particles can be filamentous virus particles. The virus particlescan possess specific binding regions on their surface at one end of thevirus particle or along the length of the viral surface. The virusparticles can also possess specific nucleation regions on their surfaceat one end of the virus particle and along the length of the viralsurface. The method can further comprise the step of blending thefibrous material with one or more other materials.

The present invention further provides a fibrous material comprising aplurality of fibers, wherein the fibers comprise one or more virusparticles which retain a viral structure in the solid state. The fibrousmaterial can further comprise non-viral blending material includingpolymer blending material, including synthetic polymer blendingmaterial. The blending material can be a water-soluble polymericblending material.

Also, the inventors have discovered a fibrous material comprising aplurality of fibers, wherein the fibers comprise one or more fiber spun,genetically engineered virus particles which retain a viral structure inthe fiber state and have specific binding sites for binding to aconjugate material. The particles can be specifically bound to theconjugate material. The particles can be capable of but not specificallybound to the conjugate material. The fibrous material can comprise atleast 50 wt. % virus particles. Or, the fibrous material can compriseless than 50 wt. % virus particles. The fibrous material can comprise atleast 50 wt. % synthetic polymer material. The fibrous material cancomprise at least two different types of virus particles. The fibrousmaterial can comprise at least one water-soluble polymer. The fibrousmaterial can comprise at least one biodegradable polymer. The fibrousmaterial can be redissolved into its viable constituent parts.

The invention further provides a fibrous material comprising aligned,crosslinked, rod-like particles, wherein the particles have a crosssectional diameter of about 5 nm to about 20 nm, and a length of about60 nm to about 6,000 nm. The length can be about 250 nm to about 1,000nm. The particles can have an aspect ratio of at least 25:1, at least75:1, or at least 100:1. The particles can be genetically engineeredvirus particles and can comprise protein and nucleic acid components.

The present invention further provides a method comprising the step of:infecting a host with a viral material, wherein the viral material isprovided from a fibrous material comprising virus particles in fiberform.

The present invention further provides a method of converting virusparticles to fiber form in which the virus particles retain their virusstructure in the solid state comprising the step of spinning the viralparticles into fiber form while controlling concentration, viscosity,and optional use of electric field and blending agent to control thefiber form and retain the virus structure after spinning. Theconcentration can be controlled so that a liquid crystalline phase canbe formed for spinning. The concentration can be sufficiently highduring spinning to avoid electrospraying.

The present invention also provides experimental conditions whichprovide the fibers with useful properties such as, for example, theability to be specifically bound to the conjugate material after fiberspinning or retain the ability to specifically bind to the conjugatematerial after fiber spinning, as well as retain the ability to infect ahost after fiber spinning. For example, concentrations can be adjusted,viscosity can be adjusted, solvents can be altered or replaced, electricfields can be applied, and use of blending materials can be varied.

In another embodiment, the present invention provides a textilecomprising a plurality of fibers, wherein each of the fibers comprises aplurality of virus particles arranged in fiber form.

In another embodiment, the present invention provides a nonwoven fabriccomprising a plurality of fibers arranged in a planar sheet, whereineach of the fibers comprises at least one virus particle arranged infiber form.

In another embodiment, the present invention provides a non-wovenmaterial or a woven material comprising fibrous material comprisingvirus particles. Also provided is a sensor comprising fibrous materialcomprising virus particles. Also provided is a biomedical devicecomprising fibrous material comprising virus particles. Also provided isa drug delivery device comprising fibrous material comprising virusparticles. The device can have fibrous material which further comprisesa conjugate material which is delivered at a controlled rate as thefibrous material dissolves upon insertion into tissues or cells. Thepresent invention also provides an article comprising a plurality offibers comprising virus particles which are genetically engineered tospecifically bind to each other.

In another embodiment, the present invention provides a garment withsensors for detecting at least one stimulus, comprising a garmentmanufactured using fibers comprising viruses engineered to express oneor more amino acid oligomers on the exterior surfaces of the virusesthat bind with a specified molecule wherein the binding causes adetectable change in the garment.

Still further, the invention provides a method of forming a viralfibrous material comprising a plurality of fibers comprising the stepsof providing virus particles and a solvent in a fiber spinningcomposition, spinning the virus particles to form the fibrous material,wherein the composition is subjected to a nucleation reaction beforespinning or the fibrous material is subjected to a nucleation reactionafter spinning to form nanocrystals in the fibrous material.

The present invention provides numerous advantages over the prior art.For example, the present invention provides a method of forming fiberson a micrometer or nanometer scale engineered for specific applications,including a wide variety of different applications ranging from militaryto biomedical. Genetic engineering can be used to engineer theapplication at an unprecedented level of structural control ranging downto the nanoscale. In another advantage, the viruses used in making thefibers of the present invention may be engineered to expresspolypeptides having one or more amino acid oligomers on the exteriorsurface of the viruses. These amino acid oligomers may serve a varietyof functions. For example, the amino acid oligomers may be cell adhesionfactors, antibodies, or have specificity for inorganic molecules. Fibersprovided by the present invention have a variety of application,including but not limited to, scaffolds for tissue engineering, sensorsfor detecting environmental toxins, reaction sites for chemicalreactions, and filters. For nanofibers, particularly when the fiberdiameter is about 100 nm or less, the materials can have exceptionallyhigh levels of specific surface area which enables a high proportion ofatoms on the fiber surface. This can provide quantum efficiency,nanoscale effect of unusually high surface energy, surface reactivity,high thermal and electrical conductivity, and high strength.

In sum, virus based micro- and nanofibers can be fabricated usingwet-spinning and electrospinning processes. M13 viruses in the wet-spunfibers were aligned parallel to the fiber long axis. Electrospun fibers,composed of fragment of M13 viruses and its subunits, were alsofabricated by suspending M13 viruses in HFP. Although the viralstructure was partially disrupted, these results indicated that thenovel biomaterials could be used to expand the dimension of theengineered viruses into endless fibers which can be used to nucleateuseful semiconductor nanofibers or to selectively bind desiredmaterials. Additionally, uniform nanofibers fabricated by blending M13virus with PVP might provide useful biological functions and highlysensitive catalytic functions in future biomedical applications andbiosensors.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows M13 virus fiber fabricated by wet-spinning processes. (A)POM image (scale bars: 100 microns), (B) SEM image (scale bars: 20microns), and (C) AFM image (scale bars: 20 microns). (D) fluorescencemicrographs of virus-phycoerythrin conjugated fibers fabricated bywet-spinning.

FIG. 2 shows (A) POM image of electrospun virus-only fibers, (B) SEMimage of electrospun virus-only fibers (scale bars: 5 microns).

FIG. 3 shows electrospun fiber of M13 virus blended with PVP. (A)Photograph of non-woven fiber spun through the mask inscribed word“NANO”, (B) SEM image (scale bar: 1 micron), (C) POM image, (D)fluorescence micrographs of PVP blended with virus-phycoerythrin fibersfabricated by electrospinning.

FIG. 4 shows a schematic diagram illustrating virus fiber fabricationprocess using wet-spinning and electrospinning.

FIG. 5 shows schematic illustration of electrospinning apparatus (A) andits photograph (B).

FIG. 6 illustrates hybrid inorganic-viral fibers and films (lower right)as well as other viral-related structures.

FIG. 7 provides characterization of wet spun virus fibers. (A) polarizedoptical microscope image (scale bar: 100 microns) (B-D) SEM images(scale bars: 100 microns (B), 20 microns (C), and (E) surface morphologyof the fiber (scale bar: one micron).

FIG. 8 provides polarized optical micrograph (left) and fluorescencemicrograph (right) of wet-spun virus conjugated with R-phycroerythrinfibers.

FIG. 9 provides optical micrograph of electrospun virus fibers (A) andits corresponding polarized optical micrograph of area drawn in dottedbox (A), scale bar (100 microns).

FIG. 10 provides SEM ranges of electrospun virus fibers and distributionof the diameter measured in SEM (scale bars: (A) 10 microns, and (B) 5microns.

FIG. 11 provides photograph of electrospun fibers of virus blended withPVP, which showed the non-woven fibers could be transformed into anyshape.

FIG. 12 provides series of SEM images of virus blended with PVP andtheir diameter distribution (D) measured in SEM, (scale bars (A), (B) 10microns and (C) one micron).

FIG. 13 provides optical micrographs of PVP blended anti-streptavidinvirus conjugated with phycoerythrin. Fluorescence micrograph of virusconjugated with phycoerythrin blended PVP (A) and a polarized opticalmicrographs of virus blended with PVP (B).

DETAILED DESCRIPTION

I. Introduction

In the present invention, virus-based micro- and nanofibers can befabricated using, for example, wet-spinning and electrospinningprocesses. The fibers demonstrate that novel biomaterials can befabricated from a programmed organism to extend the dimension ofengineered viruses into fibers useful in, for example, nucleatingsemiconductor nanofibers or in selectively binding a variety ofdifferent types of desired materials. Additionally, in one embodiment,uniform nanofibers can be fabricated by blending nanomaterial-conjugatedM13 virus with a water-soluble synthetic polymer, PVP, to provide usefulbiological functions and highly sensitive catalytic functions. Theinvention can be used for biomedical applications and biosensors.

In practice of the present invention, reference can be made to thethesis, Lee S.-W., Doctoral Thesis, The University of Texas (Austin),2003, and in particular to chapter and descriptions focused on fibers,which is incorporated herein by reference in its entirety. In addition,reference can be made to the following papers, including their figures(i) C. E. Flynn et al. Acta Materiala, 51, 5867-5880 (2003) entitled“Viruses as vehicles for growth, organization, and assembly ofmaterials.” (ii) Seung-Wuk Lee et al., Nanoletters, 4, (3), 387-390 2004entitled “Virus-Based Fabrication of Micro- and Nanofibers UsingElectrospinning,” the complete disclosures of which are incorporatedherein by reference in their entirety. In addition, priority provisionalapplication 60/510,862 filed Oct. 15, 2003 to Belcher et al. is herebyincorporated by reference in its entirety.

In addition, one skilled in the art can also refer to the followingpatent literature for selection of the virus, genetic engineeringmethods, and for materials to be used with genetically engineeredviruses: phage display libraries and experimental methods for using themin biopanning are further described, for example, in the following U.S.patent publications to Belcher et al.: (1) “Biological Control ofNanoparticle Nucleation, Shape, and Crystal Phase”; 2003/0068900published Apr. 10, 2003; (2) “Nanoscale Ordering of Hybrid MaterialsUsing Genetically Engineered Mesoscale Virus”; 2003/0073104 publishedApr. 17, 2003; (3) “Biological Control of Nanoparticles”; 2003/0113714published Jun. 19, 2003; (4) “Molecular Recognition of Materials”;2003/0148380 published Aug. 7, 2003, (5) “Composition, method, and useof bifunctional biomaterials”; 2004/0127640; filed Sep. 4, 2003; (6)“Peptide Mediated Synthesis of Metallic and Magnetic Materials”; Ser.No. 10/665,721, filed Sep. 22, 2003; and (7) “Fabricated BioFilm StorageDevice”; 2004/0171139, filed Sep. 24, 2003, which are each herebyincorporated by reference in their entirety. These references describe avariety of specific binding modifications which can be carried out forbinding to conjugate structures, as well as forming the conjugatestructures in the presence of the material modified for specificbinding. In particular, polypeptide and amino acid oligomeric sequencescan be expressed on the surfaces of viral particles, including both atthe ends and along the length of the elongated virus particle such asM13 bacteriophage, including pIII and pVIII expressions, as well as pIX,pVII, and pVI expressions, and combinations thereof.

One skilled in the art can also refer to (i) Nam et al., Nano Lett.,2003, 4, 23-27; (ii) provisional application “MULTIFUNCTIONALBIOMATERIALS AS SCAFFOLDS FOR ELECTRONIC, OPTICAL, MAGNETIC,SEMICONDUCTING, AND BIOTECHNOLOGICAL APPLICATIONS”, 60/511,102 filedOct. 15, 2003 to Belcher et al., and (iii) the regular application toBelcher et al., Ser. No. 10/965,227, “MULTIFUNCTIONAL BIOMATERIALS ASSCAFFOLDS FOR ELECTRONIC, OPTICAL, MAGNETIC, SEMICONDUCTING, ANDBIOTECHNOLOGICAL APPLICATIONS”, filed on the same day as the presentapplication, which are each incorporated by reference in their entirety,including figures, claims, and working examples.

In addition, the annealing of nanocrystalline nanowires by thermaltreatment to form annealed nanowires, and if desired, to remove theunderlying viral scaffold is described in, for example, (i) “PeptideMediated Synthesis of Metallic and Magnetic Materials”; Ser. No.10/665,721, filed Sep. 22, 2003; (ii) U.S. provisional application toBelcher et al, 60/534,102, filed Jan. 5, 2004, “Inorganic Nanowires.”;and (iii) Belcher et al., Science, 303, 213 (2004); which are eachincorporated by reference in their entirety including figures, claims,and working examples.

“Selection of Peptides with Semiconductor Binding Specificity forDirected Nanocrystal Assembly”; Whaley et al., Nature, Vol. 405, Jun. 8,2000, pages 665-668, herein incorporated by reference, shows a method ofselecting peptides with binding specificity using a combinatoriallibrary. Specifically, the article shows a method of selecting peptideswith binding specificity to semiconductor materials using acombinatorial library with about 10⁹ different peptides. Thecombinatorial library of random peptides, each containing 12 aminoacids, were fused to the pIII coat protein of M13 coliphage and exposedto crystalline semiconductor structures. Peptides that bound to thesemiconductor materials were eluted, amplified, and re-exposed to thesemiconductor materials under more stringent conditions. After the fifthround of selection, the semiconductor specific phages were isolated andsequenced to determine the binding peptide. In this manner, peptideswere selected with high binding specificity depending on thecrystallographic structure and composition of the semiconductormaterial. The technique could be readily modified to obtain peptideswith a binding specificity for not just semiconductor materials, but arange of both organic and inorganic materials.

References 4-7 noted below are also incorporated herein by reference intheir entirety. For example, Lee, S.-W.; Mao, C.; Flynn, C. E., Belcher,A. M., Science, (2002) 296, 892-895 describes methods of forminglyotropic and solid state liquid crystalline phases for hybrid viralmaterials; Lee, S.-W.; Lee, S. K.; Belcher, A. M., Adv. Mat. (2003), 15,689 further describes liquid crystalline behavior for hybrid viralmaterials; Flynn, E. C., Mao, C.; Hayhurst, A.; Williams, J. L.;Georgiou, G.; Iverson, B.; Belcher A. M., J. Mater. Chem., (2003) 13,2414 describes nucleation processes to form hybrid viral materials; andMao, Chuanbin; Flynn, Christine E.; Hayhurst, Andrew; Sweeney, Rozamond;Qi, Jifa; Georgiou, George; Iverson, Brent; Belcher, Angela M, PNAS,(2003), 100(12), 6946 also describes hybrid viral materials prepared bynucleation.

II. Virus

In the present invention, a variety of fiber structures can be made fromelongated structures. The structures can comprise biomolecules includingbiomolecular macromolecules and oligomers, including peptide portionsand nucleic acid portions. The structures can comprise naturallyoccurring materials as well as synthetic materials and geneticallyengineered materials in blended and composite arrangements. In thepreferred embodiment, these structures comprise both protein and DNAportions and, more particularly, are viruses.

For example, fibrous material can be formed comprising aligned,crosslinked, rod-like particles as fiber building blocks, wherein theparticles have a cross sectional diameter of about 5 nm to about 20 nm,and a length of about 60 nm to about 6,000 nm. More particularly, thelength can be about 250 nm to about 1,000 nm.

The virus is not particularly limited so long as fibers can be prepared.In general, virus particles which are long, filamentous structures canbe used. See, e.g., Genetically Engineered Viruses, Christopher Ring(Ed.), Bios Scientific, 2001. Virus particles which can function asflexible rods can be used.

In one embodiment, virus particles are used which are not geneticallyengineered. However, in general, desirable properties can be achievedwhen the virus is genetically engineered. In particular, viruses can beused which have been subjected to biopanning so that the virus particlesspecifically can recognize and bind to materials which were the objectof the biopanning. The viruses can be converted to fiber form with orwithout the conjugate moiety.

Use of filamentous virus in so called directed evolution or biopanningis further described in the patent literature including, for example,U.S. Pat. Nos. 5,223,409 and 5,571,698 to Ladner et al. (“DirectedEvolution of Novel Binding Proteins”).

The size and dimensions of the virus particle can be such that theparticle is elongated. For example, fibrous viral material can be formedcomprising aligned, crosslinked, rod-like particles, wherein the viralparticles have a cross sectional diameter of about 5 nm to about 20 nm,and a length of about 60 nm to about 6,000 nm. More particularly, thelength can be about 250 nm to about 1,000 nm.

Mixtures of two or more different kinds of viruses can be used. Mixturesof virus particles with non-virus materials can be used.

Virus particle can include both an entire virus and portions of a virusincluding at least the virus capsid. The term virus can refer to bothviruses and phages. Entire viruses can include a nucleic acid genome, acapsid, and may optionally include an envelope. Viruses as described inthe present invention may further include both native and heterologousamino acid oligomers, such as cell adhesion factors. The nucleic acidgenome may be either a native genome or an engineered genome. “Virusparticle” further includes portions of viruses comprising at least thecapsid.

In general, a virus particle has a native structure, wherein the peptideand nucleic acid portions of the virus are arranged in particulararrangements, which is sought to be preserved when it is spun into afiber form. The virus and/or nucleic acids may be replicated after beingfabricated into a fiber form. If during fiber formation, viralre-infectivity is lost, information may be still stored, programmed,propagated, and addressable through proteins and engineered nucleicacids, including DNA oligomers, in the viral fiber.

Viruses are preferred which have expressed amino acid oligomer asspecific binding sites. Amino acid oligomers can include any sequence ofamino acids whether native to a virus or heterologous. Amino acidoligomers may be any length and may include non-amino acid components.Oligomers having about 5 to about 100, and more particularly, about 5 toabout 30 amino acid units as specific binding site can be used.Non-amino acid components include, but are not limited to sugars,lipids, drugs, enzymes, or inorganic molecules, including electronic,semiconducting, magnetic, and optical materials.

The fibers of the present invention have varied properties depending onthe viruses comprising the fibers and the methods used to manufacturethe fibers. For the purpose of this disclosure, fiber refers togenerally cylindrical structures that may go by different names,including but not limited to, nanofibers, fibrils, nanofibrils,filaments, or nanofilments. The length, diameter, and aspect ratio(ratio of length over diameter) may vary over a considerable range.Generally, the fibers may be characterized by an aspect ratio of atleast 25, at least 50, at least 75, at least 100, or even at least 250or 500. The diameter of the fibers is typically less than about 20microns, and more particularly, less than about one micron, includingdown to about 50 nm, with a minimum diameter of the diameter of theindividual virus particles comprising the fibers. The average diametercan be, for example, about 50 nm to about 20 microns, or about 100 nm toabout 500 nm. The length of the fibers may vary considerably, buttypically will be at least 10 microns and preferably at least 50microns. Macroscopic fiber spinning with fibrous materials havinglengths on the order of mm, cm, and meters are within the invention.These macroscopic fibers can be made up of smaller fibril units as knownin the art.

A wide variety of viruses may be used to practice the present invention.The fibers may comprise a plurality of viruses of a single type or aplurality of different types of viruses. Preferably, the virus particlescomprising the fibers of the present invention are helical viruses.Examples of helical viruses include, but are not limited to, tobaccomosaic virus (TMV), phage pf1, phage fd1, CTX phage, and phage M13.These viruses are generally rod-shaped and may be rigid or flexible. Oneof skill in the art may select viruses depending on the intended use andproperties of the desired fiber.

Preferably, the viruses of the present invention have been engineered toexpress one or more peptide sequences including amino acid oligomers onthe surface of the viruses. The amino acid oligomers may be native tothe virus or heterologous sequences derived from other organisms orengineered to meet specific needs. The expression of the amino acidoligomers may serve a number of functions, including but not limited to,cell adhesion factors, trophic factors, binding sites for organic orinorganic molecules or particles or nucleation sites for organic orinorganic molecules or particles. Expression of amino acid oligomersallows the viruses and fibers comprising the fibers to be engineered tospecific applications. For example, the fibers comprising engineeredfibers may contain amino acid oligomers that initiate or enhance cellgrowth for use in tissue engineering applications and bone regeneration.In another example, amino acid oligomers with specificity for a specificinorganic molecule may be expressed to bind or nucleate the inorganicmolecule to increase the efficiency of a chemical reaction. In stillanother example, the expressed amino acid oligomer may bind and detectan organic molecule, such as a biological warfare agent. Such fiberscould be incorporated into the clothing of military personnel or firstresponders as part of a sensor system. Another embodiment is theencryption of information within the genetic code of the viruses. Theseare only a few examples of the utility of fibers made from engineeredviruses, and other applications are readily apparent to one of skill inthe art.

A number of prior art references teach the engineering of viruses toexpress amino acid oligomers and may be used to assist in practicing thepresent invention. For example, U.S. Pat. No. 5,403,484 by Ladner et aldiscloses the selection and expression of heterologous binding domainson the surface of viruses. U.S. Pat. No. 5,766,905 by Studier et aldiscloses a display vector comprising DNA encoding at least a portion ofcapsid protein followed by a cloning site for insertion of a foreign DNAsequence. The compositions described are useful in producing a virusdisplaying a protein or peptide of interest. U.S. Pat. No. 5,885,808 bySpooner et al discloses an adenovirus and method of modifying anadenovirus with a modified cell-binding moiety. U.S. Pat. No. 6,261,554by Valerio et al shows an engineered gene delivery vehicle comprising agene of interest and a viral capside or envelope carrying a member of aspecific binding pair. U.S. Published Patent Application 2001/0019820 byLi shows viruses engineered to express ligands on their surfaces for thedetection of molecules, such as polypeptides, cells, receptors, andchannel proteins.

III. Conjugate

In the present invention, the virus can be conjugated with a conjugatematerial before fiber spinning or after fiber spinning.

The conjugate material is not particularly limited. In general, it willbe selected for a particular application. The conjugate material can beconjugated to the virus particles by being subjected to viral biopanningagainst the conjugate material, and then the conjugate material isspecifically bound to the virus particle and the fiber formed.Alternatively, fibers can be formed and then the conjugation carried outin, for example, a surface treatment. Conjugate material can bepreformed and then bound to the virus or it can be directly formed ornucleated on the virus. The virus can act as a catalyst for formation ofor biomineralization of the conjugate material on the virus.

Examples of general types of conjugate materials include inorganic,organic, particulate, nanoparticulate, small molecule, singlecrystalline, polycrystalline, amorphous, metallic, magnetic,semiconductor, polymeric, block copolymer, functional polymer,conducting polymeric, light-emitting, phosphorescent, organic magnet,chromophore, protein, peptide, nucleic acid, DNA, RNA, oligonucleotide,drugs, enzymes, and fluorescent materials. Conjugate materials aredescribed further, for example, in the patent publications and technicalliterature to Angela Belcher and co-workers cited in this specification.

The conjugate material can be directly bound to the virus, or can belinked to the virus by an intermediate linking moiety which can bothbind to the virus and the conjugate material.

In a preferred embodiment, the conjugate material is a nanomaterial.Examples include semiconductor nanocrystals or quantum dots which haveaverage diameters small enough to provide quantum confinement effects.Average diameters can be about 10 nm or less, or more particularly,about 5 nm or less. Nanowires can also be formed by conjugation ofnanocrystalline materials along the length of the virus and annealingthe nanocrystalline materials into nanowires. The crystals can be singlecrystals and can be oriented in the same direction.

IV. Fiber Spinning Including Wet Spinning

Many methods can be used to convert the virus to fiber form, whetheralone or with one or more blending materials. The invention encompassesa variety of fibrous structures including microfibers and nanofibers.See, e.g., Spinning, Extruding, and Processing of Fibers: RecentAdvances, J. S. Robinson, Noyes, 1980; and Advanced Fiber SpinningTechnology, Toshinari Nakajima (Ed.), Woodhead Pub., 1994. Spinninggenerally can be used in fiber production to extrude a solution throughan orifice and, if desired, subsequent cross-linking to form longerfibers. Fibers can be spun which have aspect ratios of 50 or more, 100or more, 500 or more, and 1,000 or more.

Fiber spinning is generally described in Textbook of Polymer Science,3^(rd) Ed., F. Billmeyer, 1984, Chapter 18, pages 486-505, includingtextile and fabric properties, spinning, fiber after-treatments, andfiber properties, which is hereby incorporated by reference in itsentirety. After-treatments include steps taken before weaving includingwashing, scouring, sizing, lubricating, dying, and property adjustmentsfor control of crease resistance, softness, water repellency, slipping,dimensional stability, shrinkage, and the like. In non-woven fabrics,fibers can be bonded together into flat sheets by heat, pressure, andbonding agents. Finishes can be applied to the fibers. Conventionalmethods in the art can be used for making non-woven fabrics. Fibers andfiber spinning are also described in the Concise, Encyclopedia ofPolymer Science and Engineering, J. Kroschwitz, Ed. 1990, “Fibers”,pages 374-395.

The present invention teaches methods of forming a fiber comprising thesteps of providing a solution or suspension comprising a plurality ofvirus particles and a solvent and spinning the solution or suspension.The virus particles in solution may be selected based on a variety ofcriteria as described previously. The solvent selected will depend onthe choice of virus particle. Often, the solvent will simply be waterwith buffering agents, such as TRIS-HCl. The solution may includeadditional components. Such additional components include, but are notlimited to, viscosity modifying agents, ionic strength modifying agents,aligning agents, and buffering agents. Spinning processes are well-knownin the fiber manufacturing art and include wet spinning, dry spinning,and electrospinning. Spinning processes can be selected based on thespecific application of the resulting fibers. In general, compositionsfor spinning are selected to avoid components in amounts sufficientlylarge to interfere with the fiber forming process.

In a preferred embodiment of the present invention, the solutioncomprising viruses and a solvent will be a liquid crystal solutionincluding lyotropic solutions. Fibers formed from liquid crystalsolutions have been shown to have desirable physical properties, such ashigh strength and uniformity. “Model Study of the Spinning ofThermotropic Liquid Crystalline Polymers: Fiber Performance predictionsand Bounds on Throughput”; Forest et al., Advances in PolymerTechnology, vol. 18, No. 4, pages 314-35 provides background informationon the spinning of liquid crystalline solutions and is hereinincorporated by reference.

A number of viruses have been shown to form liquid crystal phases insolution. Typically, the viruses are generally rod-shaped and may berigid or flexible. The structure can have a portion which is rigid, andthe rigid portions are connected by flexible portions, so that the virusparticle overall is semirigid. Examples of viruses forming liquidcrystal phases in solution include, but are not limited to, tobaccomosaic virus (TMV), phage pf1, phage fd, and phage M13. The phases ofthe liquid crystalline viruses may be altered by changing the solutionconcentrations, ionic strength of the solution, or altering an externalmagnetic field or flow velocity. Methods of changing the phase will bereadily apparent to one of skill in the art. The following documentsgive background information generally regarding liquid crystal virussolutions and phase adjustments that may be used to practice the presentinvention and are herein incorporated by reference: (1) “Smectic Phasein a Colloidal Suspension of Semiflexible Virus Particles”; Dogic etal., Physical Review Letters, Vol. 78, No. 12, pages 2417-20 and“Ordering of Quantum Dots Using Genetically Engineered Viruses”; Lee etal., Science, Vol. 296, pages 892-95. It is not necessary for thesolutions containing viruses to be in a particular phase in order tospin fibers.

The spinning processes that may be used in the present invention arewell-known within the art and include, but are not limited to, dryspinning, wet spinning, and electrospinning. In dry spinning, thesolution comprising virus particles and a solvent is passed through aspinneret into a zone where the solvent is quickly evaporated leavingfibers that may be wound. Fibers produced by wet spinning are formed bypassing the solution comprising virus particles and solvent through aspinneret into a nonsolvent that coagulates the virus particles. Achemical reaction may be involved in forming fibers using this process.Crosslinking can occur. The specific parameters of the spinningprocesses may be readily determined by one of ordinary skill in the art.

Preferred embodiments include wet fiber spinning, electrofiber spinning,and lyotropic liquid crystalline fiber spinning.

Wet spinning is known in the art including use of wet spinning to formfibrous, microfibrous, and nanofibrous materials.

The solvents and suspension agents used are not particularly limited.The wet spinning can be carried out, for example, with use of aqueoussolutions and suspensions. Concentrations can be used so that lyotropicliquid crystalline solutions can be formed including—smectic and nematicsolutions. Concentrations can be, for example, about 10 to about 1,000mg/mL, and more particularly, about 50 mg/ML to about 250 mg/mL. pH canbe adjusted as needed.

Capillary tubes can be used for extrusion. The size can be, for example,about 1 micron to about 100 microns, and more particularly, about 5microns to about 50 microns.

Crosslinking of the fibrous structures can be carried out. Thecrosslinking agent is not particularly limited but can be electrophilicagents, including carbonyl moieties such as, for example, glutaraldehydesolutions. Concentration of the crosslinking agent can be, for example,about 10% to about 60%, and more particularly, about 25% to about 50%.

When microfibers are made, the diameters of the microfibers can be, forexample, about one micron to about 100 microns, and more particularly,about 5 microns to about 50 microns, and more particularly about 10microns to about 20 microns.

Experimental variables such as crosslinking agent, concentration,extrusion rate, and size of capillary can be adjusted to provide thedesired fibers.

Wet spinning is illustrated in FIG. 4.

V. Electrospinning

Electrospinning of fibers in known in the art and is described in thepatent literature at, for example, U.S. Pat. Nos. 6,106,913 and6,308,509 to Scandino et al. (“Fibrous Structures Containing Nanofibrilsand Other Textile Fibers”); U.S. Pat. No. 6,110,590 to Zarkoob et al.(“Nanofibers and a Process for Making the Same”); U.S. Pat. Nos.6,382,526 and 6,520,425 to Reneker et al. (“Process and Apparatus forthe Production of Nanofibers”); and U.S. Pat. No. 6,616,435 to Lee etal. (“Apparatus of Polymer Web by Electrospinning Process”). Inaddition, the following references including background material onelectrospinning techniques may be used to practice the present inventionand are herein incorporated by reference: (1) “Nanostructured Fibers viaElectrospinning”; Bognitzki et al., Adv. Mater., Vol. 13, No. 70, pages70-XX (2001) and “Preparation of Fibers with Nanoscaled Morphologies:Electrospinning of Polymer Blends”; Bognitzki et al., Polym. Eng. Sci.,in press (2001).

Electrospinning is also illustrated in FIGS. 4 and 5 (includingphotograph of apparatus).

Volatile organic solvents can be used for electrospinning. Conditionsfor suspending the virus in the solvent can be adjusted to provide thedesired fibers. Solvents are preferred which provide a homogeneous virussuspension. Also, the solvent should allow the virus to retain itsability to infect. Concentrations can be used so that lyotropic liquidcrystalline solutions can be formed including smectic and nematicsolutions. Concentrations can be, for example, about 10 to about 1,000mg/mL, and more particularly, about 50 mg/ML to about 250 mg/mL.

Spinning conditions can be also adjusted including, for example, pumpfeed rate, applied potential, collection plate, and distance betweencapillary and collector plate. Other parameters include polymer andviral concentration, molecular weight, solution viscosity, Berry Number,boiling point, distance, voltage, and orifice diameter.

VI. Blending Material

The fiber spinning of the virus particles can be carried out with anon-viral blending material for the virus particles, including apolymeric blending material. This can also be a carrier. The blendingmaterial can be a synthetic polymer blending material. It can be awater-soluble polymeric blending material. It can be, for example, avinyl polymer which has polar side groups which can, if desired, providefor water solubility. Examples include polycarbonate, DNA, silk, aramid,polystyrene, polybutadiene, nylon, polyester, PLGA, PLAGA,polyacrylonitrile, polyaniline and other conjugated polymers, and PVDF.A preferred example is PVP. Polymers can be selected which are glassy atroom temperature, having a glass transition temperature above 25° C. Themolecular weight of the blending material can be selected to be highenough to provide for good fiber properties. For example, molecularweight (Mn) of at least 100,000, or of at least 500,000, or of at least1,000,000 can be selected.

This infection ability of the M13 virus fibers blended with PVP can beuseful in biomedical applications and tissue engineering. One skilled inthe art can tailor the infectability to the level desired for thatapplication. In some applications, infectability may be undesired. Insome applications, mere storage of the virus, including informationstored in the DNA, may be useful without need for the infectability. Inother applications, the high levels of infectability may be an importantparameter.

The relative concentrations of the virus and the blending material canbe adjusted to provide the desired fibral structures. In someembodiments, the blending material is more than 50 wt. % of the fiber,whereas in other embodiments, the blending material is less than 50 wt.% of the fiber.

The fiber diameter can be, for example, about 10 nm to about 1,000 nm,or more particularly, about 25 nm to about 500 nm, and moreparticularly, about 50 nm to about 250 nm.

Fibers can be further annealed and oriented as desired.

VII. Applications

The fibers can be arranged into desired shapes using masks andpatterning. Fiber processing methods can be used including yarnprocessing.

Applications of the invention are many and include

1) Diagnostics (medical or environmental): filter material that isoptimized for aqueous samples (because viruses are intrinsically waterloving) that contain specific binding sites useful in flow throughdiagnostic assays (e.g. sandwich assays Ab-Ag-Ag-label, or competitionassays where a labeled ligand is displaced). Flow through could be inair. Subsequent labeling could occur in liquid.

2) Diagnostic applications in (1) above can be extended by usingmultiple virus types for multiple analyte tests, thus producing a test“cocktail” in one membrane material.

3) Bioreactors (therapeutics or other products): fiber meshes or fiberbundles may be used in flow-thru bioreactors where virus components aredesigned to bind to certain cells and/or contact cells with stimulatingagents (e.g. cytokines), Viral conjugate components or viralcrosslinking may be designed to release cells or stimulate cells onlyafter a chemical change is made in the flowing media.

4) Non-woven fabrics comprising fibers or fibrous materials withspecific optical, electrical, or magnetic properties that areincorporated into textiles as sensors, detectors, and monitors. Militaryapplications can be carried out using soldier gear and uniforms whichhave the viral fibers.

5) membranes

6) sensors

Viral fibers are further illustrated in FIG. 6, as well as filmstructures. Other viral structures useful in many applications are shown(see Belcher patent publications and other paper publications citedherein).

If desired, the fibers can be heat treated and annealed. If desired,organic aspects of the fibers can be removed by thermal treatments athigh enough temperatures so that the organic component is “burned off”to leave an inorganic conjugate component, such as a nanowire, which isstable to the high temperatures. In addition, fibers can be modified andgenetically engineered so that they can recognize and couple to eachother at their ends, or along the sides.

The fibers can be attached to pre-patterned surfaces to form patterns offibers on the surface and can be deposited in patterned arrangements onsurfaces.

Viral fibers can be blended with non-viral fibers.

WORKING EXAMPLES

The invention is further described with use of the followingnon-limiting working examples.

A. Virus

Anti-streptavidin M13 bacteriophage possessing an engineered peptidesequence, N′-TRP ASP PRO TYR SER HIS LEU LEU GLN HIS PRO GLN-C′ (SEQ IDNO.:1), in its pIII coat protein (virus) was used as a basic buildingblock to fabricate the micro- and nanoscale fibers. The virus wasselected from the PhD-12 phage display library (New England Biolabs,Inc. Beverly, Mass.) for affinity to streptavidin (5). The virus wasamplified and purified according to phage library manufacturerinstructions and suspended in tris buffered saline (TBS; 50 mM Tis, 150mM NaCl, pH 7.5).

B. Conjugate Material.

The bacteriophage was used with or without conjugation withR-phycoerythrin (eBioscience, CA) previously reported [5]. Conjugationwas at the pIII subunit.

C. Wet Spinning

The M13 virus suspension (˜100 mg/ml) extruded through ˜20 um capillarytube into 37.3% aqueous glutaraldehyde solution, were cross-linked andformed microfibers. The fibers were taken and dried in the air.Polarized optical microscopy image (FIG. 1A) taken using Olympus IX51polarized optical microscope (POM; Olympus, Japan) under the crossedpolars exhibited birefringent, which indicates liquid crystallineordered structures of the fibers. SEM images taken using a JEOL 6320FEGSEM field emission scanning electron microscope (FE-SEM; JEOL, Japan)(FIG. 1B) of the fibers showed that the fibers had 10-20 micrometer indiameter and were composed of several bundle-like fibers which werepropagated to fiber long axis. Parallel orientation of the individualstand of virus building block was observed by AFM. AFM image (FIG. 1C)showed close-packed M13 virus of which long axis was parallel to thelong axis of the fibers. Although smectic suspension was spun tofabricate the virus fibers, smectic layered structures were not observedfrom POM and AFM. Due to flowing field, smectic ordered structure in thesuspension was disrupted and smectic to nematic transitions occurred.Because the suspension was immediately exposed to cross-linking solutionafter releasing from the capillary, the nematic ordered viruses werecross-linked each other and formed nematic ordered fibers. Florescentviral microfibers were fabricated by spinning fibers after conjugationof virus and R-phycoerythrin (eBioscience, CA) covalently conjugatedwith streptavidin [5]. Uniform fluorescent light (FIG. 1D) throughoutthe fibers supported the nematic ordered structures observed in POM andAFM which did not have positional ordered structures.

Wet spun fibers are further shown in FIGS. 7 and 8.

D. Electrospinning

Aqueous M13 virus suspension could not be electrospun butelectro-sprayed because the suspension exhibited too low viscosity to bespun. However, M13 viruses suspended in a volatile organic solvent couldbe spun using electrospinning. Various organic solvents were tested forthe possibility as a spinning solvent. However, most of theelectrospinning solvents tested, such as methanol, ethanol, DMF, acetoneand trifluoroethanol, formed slurry-like aggregation when they wereadded to virus pellets and suspensions.1,1,1,3,3,3,hexafluoro-2-propanol (HFP; Alpha Aeser, MA) was the bestsolvent to form homogenous virus suspension when the virus pellets weredissolved. In order to fabricate nanofibers, capillary tubes (20 ulvolume, ˜0.5 mm in diameter, 6.5 cm long, Drummond Scientific Co., PA)were filled with virus suspension (˜92 mg/ml in HFP), and a graphite rod(0.5 mm in diameter) was inserted into one end of the capillary tube bya syringe pump (Harvard Apparatus, Inc., MA) at a feeding rate of 3-6ul/min. A 20-30 kV potential was applied by a high voltage source acrossair (Glassman High Voltage, NJ). Electrospun fibers were collected ontometal plates (10 cm in diameter) grounded and covered by aluminum foil.Distance between capillary and collector was 10-15 centimeters.Polarized optical microscopy image (FIG. 2A) verified that thesecontinuous electrospun fibers were highly birefringent under the crosspolars, indicating that there were highly crystalline ordered structuresin the fibers. Relatively broad distribution of diameter was observedranging from a few micrometers to tens of nanometers in diameter. SEMimage (FIG. 2B). shows that most of the fibers have branched shapedseparated from larger fiber bundles to smaller fibers. Due to thetoxicity of HFP to the M13 virus, infectability of M13 virus in HFPsolution was dramatically decreased and showed no infectability. Becauseintact virus structures were rarely observed from the TEM observation,virus fibers spun using HFP could be composed of the fragment of virusand dissembled subunits.

Electrospun fibers are further shown in FIGS. 9 and 10.

E. Electrospinning with Blending Material

In order to improve processing ability and preserve the intact viralstructure and infecting ability, M13 virus suspension was blended withhighly water soluble polymer, polyvinyl 2-pyrolidone (PVP; M.W.:1,300,000; (Alfa Aesar, MA). The suspensions blended in ratio of 1:1,1:2, 1:3 and 1:4 between virus suspension in TBS (˜100 mg/ml) and PVPsolution (25%, (w/w)) in water. Due to the low viscosity and highsurface tension of the aqueous suspensions, 1:1 and 1:2 suspensions werenot electrospun but deposited droplets of virus-PVP suspension. The 1:3suspension was sporadically spun and formed bead and string type fibers,which is normally observed in low viscous or concentration solutions inelectrospinning process [29-31]. However, there were still manyelectrosprayed droplets observed. Continuous M13 virus blended PVPfibers were fabricated from the 1:4 suspension. Photographs ofelectrospun fibers (FIG. 3A) showed that electrospun fibers could betransformed to any shape of non-woven fabrics. A mask was made fromsimple paper cut-outs placed over a glass substrate on the collectorplate. SEM image showed that the resulting fibers were continuous andhomogeneous round rope shapes (FIG. 3B). Due to the lack of drivingforce for the orientation of the fibers, such as rotating mandrels[32,33] or a conducting-gap mounted collector [26], no preferredorientation of the fibers was observed. Distribution of the diameter wasrelatively narrow although a few micrometers were, still observed. Mostof the fibers had diameter ranging 100-200 nanometers. The electrospunfibers observed using polarized optical microscope exhibitednematic-like birefringent (FIG. 3C). The fibers showed maximumbrightness when the fibers were oriented ˜45 degrees to the cross polarsand extinct when the fibers were oriented to parallel with cross polars.When the viruses conjugated with R-phycoerythrin using streptavidinlinker, fluorescence images could be observed using fluorescencemicroscope (FIG. 3D). After dissolving the electrospun fibers in 3 ml oftris-buffered saline (pH: 7.5), the suspension was taken to testinfecting ability of the M13 virus. The resuspended virus suspensionshowed that the virus still active to infect the bacterial host.

FIGS. 11-13 further show data from the PVP viral fiber system.

The non-woven fibers can be deposited in higher concentrations in orderto synthesize self-supporting non-woven fabrics. Substrates used duringthe deposition of the non-woven fiber fabrics can be further optimizedto further induce self-supporting and oriented systems.

One skilled in the art can further practice the present invention byreference to the following references, which are hereby incorporated byreference in their entirety.

REFERENCES

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1. A method of forming a viral fibrous material comprising a pluralityof fibers and a non-viral blending material comprising providing virusparticles, a non-viral blending material, and a solvent in a fiberspinning composition, and then spinning the virus particles to form thefibrous material.
 2. The method according to claim 1, wherein thespinning is an electrospinning.
 3. The method according to claim 1,wherein the virus particles are filamentous virus particles.
 4. Themethod according to claim 1, wherein the virus particles possessselective binding regions on their surface and are bound to a conjugatematerial.
 5. The method according to claim 4, wherein the conjugatematerial is an inorganic or an organic material.
 6. The method accordingto claim 1, wherein the fiber spinning is carried out with crosslinkingof the viral particles to form cross-linked fiber.
 7. The methodaccording to claim 1, wherein after spinning the fibrous material isliquid crystalline.
 8. A method of forming a genetically engineeredfibrous material comprising a plurality of fibers and a non-viralblending material comprising the step of fiber spinning geneticallyengineered virus particles with a non-viral blending material to formthe fibrous material, wherein the virus particles of the fibrousmaterial are specifically bound to a conjugate material after fiberspinning or are capable of specifically binding to a conjugate materialafter fiber spinning, and the virus particles of the fibrous materialretain the virus structure after fiber spinning.
 9. A fibrous materialcomprising a plurality of fibers, wherein the fibers comprise one ormore fiber spun, genetically engineered virus particles which retain aviral structure in the fiber state and have specific binding sites forbinding to a conjugate material, wherein the virus particles compriseexpressed oligopeptide sequences which provide the specific binding. 10.The fibrous material according to claim 9, wherein the conjugatematerial is a semiconductor material, a magnetic material, or a metallicmaterial.
 11. The fibrous material according to claim 9, wherein theconjugate material is a crystalline material.
 12. The fibrous materialaccording to claim 9, wherein the conjugate material is a drug orenzyme.
 13. The fibrous material according to claim 9, wherein the virusparticles have specific binding sites at their ends, and along theirlengths.
 14. The fibrous materials according to claim 9, wherein thefibrous material further comprises metallic, ceramic, or glassymaterial.
 15. The fibrous materials according to claim 9, wherein thefibrous material further comprises polymeric material.
 16. The fibrousmaterial according to claim 9, wherein the fibrous material comprises atleast two different types of virus particles.
 17. The fibrous materialaccording to claim 9, wherein the virus particles are filamentous virusparticles and the fibrous material further comprises a blendingmaterial.
 18. The fibrous material according to claim 17, wherein thefibrous material can be redissolved into its viable constituent parts.19. A method comprising the step of: infecting a host with a viralmaterial, wherein the viral material is provided from a fibrous materialcomprising virus particles and a non-viral blending material in fiberform.
 20. A method of converting virus particles to fiber form in whichthe virus particles retain their virus structure in the solid statecomprising the step of spinning the viral particles into fiber form witha non-viral blending material while controlling concentration,viscosity, and optional use of electric field to control the fiber formand retain the virus structure after spinning.
 21. A fibrous materialcomprising a plurality of fibers, wherein the fibers comprise one ormore fiber spun, genetically engineered virus particles which retain aviral structure in the fiber state and have specific binding sites forbinding to a conjugate material, wherein the conjugate material is asemiconductor material, a magnetic material, or a metallic material. 22.A fibrous material comprising a plurality of fibers, wherein the fiberscomprise one or more fiber spun, genetically engineered virus particleswhich retain a viral structure in the fiber state and have specificbinding sites for binding to a conjugate material, wherein the conjugatematerial is a crystalline material.
 23. A fibrous material comprising aplurality of fibers, wherein the fibers comprise one or more fiber spun,genetically engineered virus particles which retain a viral structure inthe fiber state and have specific binding sites for binding to aconjugate material, wherein the conjugate material is a drug or enzyme.24. A fibrous material comprising a plurality of fibers, wherein thefibers comprise one or more fiber spun, genetically engineered virusparticles which retain a viral structure in the fiber state and havespecific binding sites for binding to a conjugate material, wherein thefibrous material further comprises polymeric material.
 25. A wovenmaterial comprising the fibrous material of claim
 9. 26. A sensorcomprising the fibrous material of claim
 9. 27. A biomedical devicecomprising the fibrous material of claim
 9. 28. A drug delivery devicecomprising the fibrous material of claim 9.